tag:blogger.com,1999:blog-9031065214003247297.post207346864112466042..comments2016-02-07T13:32:05.041-08:00Jake Hearenoreply@blogger.comBlogger1125tag:blogger.com,1999:blog-9031065214003247297.post-32772217561678778912015-05-04T22:18:43.528-07:002015-05-04T22:18:43.528-07:00A few notes:<br /><br />- Probably not a big deal, but you should&#39;ve used 22uL of DNase Inactivation Reagent (protocol says to use 0.2 volumes if you used 2uL of DNase).<br /><br />- You should specify which DNA sample you used for your positive control (e.g. label, date isolated, concentration)<br /><br />- Your qPCR reaction doesn&#39;t add up. The Sso Fast is a 2x master mix. So, if you&#39;re using 10uL of Sso Fast, your final reaction volume (including template) should be 20uL. In this instance, it&#39;s not too big of deal, but for future reference this could impact your reactions if you deviate too far from the appropriate dilution.<br /><br />- You mentioned a negative control, but you didn&#39;t actually run a negative control. To clarify, a negative control reaction would receive DNA template that you know will not produce amplification. Then, there&#39;s a no template control, in which you substitute water for template in the reaction. In your case, you actually don&#39;t have either because the reaction you label as a negative control isn&#39;t the same volume (because it&#39;s lacking template or water in place of template) as all the other reactions on your plate.<br /><br />- You need to add your qPCR plate layout to your notebook.<br /><br />- You need to add a link to the raw qPCR data file to your notebook, not a CSV export. The raw datafile is of file type TAD.Sambhttp://www.blogger.com/profile/03618493573679963503noreply@blogger.com